Type II (immune) interferon preparations have been shown to possess the important biological properties of being antiviral, antitumor, and immunoregulatory. Current schemes for production of immune interferon are inadequate for purification in order to study the above biological activities in an efficient manner. We propose to produce mouse immune interferon (type II) in a large-scale primary culture system. The produced interferon will be purified by the following types of chromatography: (1) affinity hydrophobic columns such as phenyl-Sepharose and BSA-Affi-Gel 10; (2) adsorption chromatography on Control Pore Glass beads; (3) gel filtration columns such as Ultrogel AcA 34 and AcA 54. It is estimated that we can produce 1-2 x 10 to the 7th power units of immune interferon/week in a modified roller culture system using C57B1/6 mouse spleen cells and the T cell mitogen staphylococcal enterotoxin A as the interferon inducer. Preliminary data where the above columns have been used in combination has resulted in approximately 1,000 - fold purification of immune interferon with yields of 80 - 90% of starting material. The use of chromatography in combination resulted in immune interferon with greater than 10 to the 6th power units/mg protein, free of MIF and lymphotoxin activities, but possessing immunosuppressive activity. The purification scheme presented here will allow further purification in conjunction with the identification of the antiviral, antitumor, and immunosuppressive activities present in immune interferon preparations. Antisera will be produced to the immune interferon. This study, although not designed to purify MIF and lymphotoxin, will monitor these lymphokine activities in the immune interferon purification as a part of the characterization of the purified interferon. It is hoped that this virus study will definitively determine the biological properties of the characterized type II interferon.